Population dynamics of rumen microbes using modern techniques in rumen enhanced solid incubation

2003 ◽  
Vol 48 (4) ◽  
pp. 113-119 ◽  
Author(s):  
N. Raizada ◽  
V. Sonakya ◽  
R. Dalhoff ◽  
M. Hausner ◽  
P.A. Wilderer

The microbial ecology of the rumen is very complex. Different species of bacteria, protozoa, and fungi are involved in digestion of plant material in ruminants. In spite of complicated interrelationships among the various groups of microorganisms in the rumen ecosystem, Bacteria and Archaea are believed to play a major role because of their numerical predominance and metabolic diversity. In this work we are presenting the results for microbial population dynamics of rumen microbes during two-stage anaerobic digestion of grass. The reactors were inoculated with fresh rumen content. Fluorescent in situ hybridization, confocal laser scanning microscopy and epifluorescence microscopy were employed for microbial investigation. It was observed that Bacteria dominated in the hydrolytic reactor (1st stage) whereas Archaea were predominant in the methanogenic reactor (2nd stage). The stability of the methanogenic reactor was result of the dominance of Methanosaeta species (mainly the filamentous type).

Author(s):  
Thomas M. Jovin ◽  
Michel Robert-Nicoud ◽  
Donna J. Arndt-Jovin ◽  
Thorsten Schormann

Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.


Materials ◽  
2021 ◽  
Vol 14 (8) ◽  
pp. 1821
Author(s):  
Ting He ◽  
Wei Shi ◽  
Song Xiang ◽  
Chaowen Huang ◽  
Ronald G. Ballinger

The influence of AlFeSi and Mg2Si phases on corrosion behaviour of the cast 6061 aluminium alloy was investigated. Scanning Kelvin probe force microscopy (SKPFM), electron probe microanalysis (EPMA), and in situ observations by confocal laser scanning microscopy (CLSM) were used. It was found that Mg2Si phases were anodic relative to the matrix and dissolved preferentially without significantly affecting corrosion propagation. The AlFeSi phases’ influence on 6061 aluminium alloy local corrosion was greater than that of the Mg2Si phases. The corroded region width reached five times that of the AlFeSi phase, and the accelerating effect was terminated as the AlFeSi dissolved.


Life ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 301
Author(s):  
Kathrin Malfeld ◽  
Nina Armbrecht ◽  
Holger A. Volk ◽  
Thomas Lenarz ◽  
Verena Scheper

In recent years sensorineural hearing loss was found to affect not exclusively, nor at first, the sensory cells of the inner ear. The sensory cells’ synapses and subsequent neurites are initially damaged. Auditory synaptopathies also play an important role in cochlear implant (CI) care, as they can lead to a loss of physiological hearing in patients with residual hearing. These auditory synaptopathies and in general the cascades of hearing pathologies have been in the focus of research in recent years with the aim to develop more targeted and individually tailored therapeutics. In the current study, a method to examine implanted inner ears of guinea pigs was developed to examine the synapse level. For this purpose, the cochlea is made transparent and scanned with the implant in situ using confocal laser scanning microscopy. Three different preparation methods were compared to enable both an overview image of the cochlea for assessing the CI position and images of the synapses on the same specimen. The best results were achieved by dissection of the bony capsule of the cochlea.


2001 ◽  
Vol 67 (11) ◽  
pp. 5273-5284 ◽  
Author(s):  
Holger Daims ◽  
Jeppe L. Nielsen ◽  
Per H. Nielsen ◽  
Karl-Heinz Schleifer ◽  
Michael Wagner

ABSTRACT Uncultivated Nitrospira-like bacteria in different biofilm and activated-sludge samples were investigated by cultivation-independent molecular approaches. Initially, the phylogenetic affiliation of Nitrospira-like bacteria in a nitrifying biofilm was determined by 16S rRNA gene sequence analysis. Subsequently, a phylogenetic consensus tree of theNitrospira phylum including all publicly available sequences was constructed. This analysis revealed that the genusNitrospira consists of at least four distinct sublineages. Based on these data, two 16S rRNA-directed oligonucleotide probes specific for the phylum and genus Nitrospira, respectively, were developed and evaluated for suitability for fluorescence in situ hybridization (FISH). The probes were used to investigate the in situ architecture of cell aggregates ofNitrospira-like nitrite oxidizers in wastewater treatment plants by FISH, confocal laser scanning microscopy, and computer-aided three-dimensional visualization. Cavities and a network of cell-free channels inside the Nitrospiramicrocolonies were detected that were water permeable, as demonstrated by fluorescein staining. The uptake of different carbon sources byNitrospira-like bacteria within their natural habitat under different incubation conditions was studied by combined FISH and microautoradiography. Under aerobic conditions, theNitrospira-like bacteria in bioreactor samples took up inorganic carbon (as HCO3 − or as CO2) and pyruvate but not acetate, butyrate, and propionate, suggesting that these bacteria can grow mixotrophically in the presence of pyruvate. In contrast, no uptake by theNitrospira-like bacteria of any of the carbon sources tested was observed under anoxic or anaerobic conditions.


2000 ◽  
Vol 41 (12) ◽  
pp. 69-77 ◽  
Author(s):  
J. C. Araujo ◽  
G. Brucha ◽  
J. R. Campos ◽  
R. F. Vazoller

In this study we investigated the development of anaerobic biofilm using a laboratory reactor. We were especially interested in comparing the organization of anaerobic cells (particularly those that are very common in domestic sewage sludge) in a hydrophilic (glass) versus a hydrophobic (polypropylene) surface. Fluorescent in situ hybridization (FISH) with domain and group specific probes directed against 16S ribosomal RNA were used to quantify microbial composition in the biofilm. FISH and confocal laser scanning microscopy (CLSM) were used to elucidate spatial distribution of microbes in the biofilms. Two experiments were carried out, one with pure methanogenic organisms and the other with a microbial anaerobic consortium. The pure methanogen cultures, Methanobacterium formicicum (DSM 1535); Methanosaeta concilli (DSM 3671) and Methanosarcina barkeri (DSM 800) were used to seed the modified Robbins Device (MRD) to allow the development of biofilms on polypropylene and glass surfaces during the 9-days experiment. The results showed that all the three species were colonizing both surfaces after two and nine days of experimental period. In another experiment, with polypropylene coupons only, MRD was seeded with a microbial anaerobic consortium and biofilm formation was studied during 11 days. At the end of this period, the biofilms generated were of uneven thickness with areas of minimal or no surface coverage and areas where the biofilm attained a thickness of 7.0 to 9.0 μm as revealed by CLSM. The results showed that the modified Robbins Device together with the fluorescent in situ hybridization and confocal laser scanning microscopy are suitable tools to study anaerobic biofilm development in different kinds of support materials.


2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Hiroyuki Yamamoto ◽  
Hiroki Sakagami ◽  
Yoshio Kijidani ◽  
Junji Matsumura

A modified confocal laser scanning microscopy (CLSM) system was developed not only to observe the microcracks on the surface ofCryptomeria japonicaD. Donin situat the cellular level but also to obtain information about the moisture content (MC) of the wood surface by measuring the change in its electrical resistivity. The sequential images and changes in the electrical resistivity of the wood surface indicated that microcracks formed between the tracheid and ray parenchyma in the latewood region at >1.0E+ 07 Ω/sq (square). Microcracks formed when the MC of the wood surface was below the fiber saturation point determined through regression analysis of the surface electrical resistivity and MC. Most of the microcracks develop when the surface electrical resistivity ranged from 3.95E+ 10 to 3.60E+ 12 Ω/sq. When the surface MC was~2.5%, microcracks closed and the surface electrical resistivity was either~1.00E+ 15 Ω/sq or outside the measurement range. The modified CLSM and the method to measure the MC of the wood surface can be used to acquire information about the surface MC in specific areas shown in CLSM images. The findings indicated that the MC of the surface of the wood plays an important role in suppressing the emergence of microcracks in drying wood. The modified CLSM system and the method of measuring the MC of the surface of wood can be used to efficiently evaluate methods of drying wood and the quality of dried wood.


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